OPTIMIZED DIFFERENTIAL DISPLAY AND REAMPLIFICATION PARAMETERS FOR SILVER STAINING

Differential display (DD) is a powerful instrument to detect differenc es in gene expression of malignant and normal cells. An optimized silv er staining protocol was used to compare mRNA expression of keratinocy tes and squamous cell carcinoma cells of the head and neck. Optimal co ncentration for upstream and downstream primer was 0.2 mu M. Best prim er concentrations for reamplification were between 40 and 60 pM. With this optimized protocol nearly 50 differentially expressed transcripts have been identified and differential display can be applied safer, e asier, and faster, than by radioactive labeling.